Binary mixtures of feeding deterrents mitigate the decrease in feeding deterrent response to antifeedants following prolonged exposure in the cabbage looper, <Emphasis Type="Italic">Trichoplusia ni</Emphasis> (Lepidoptera: Noctuidae)

Summary. The effect of rearing larvae of Trichoplusia ni on individual feeding deterrents or on binary mixtures of deterrents on their subsequent gustatory sensitivity was measured in paired choice leaf disc bioassays. Our working hypothesis was that mixtures of antifeedants (pure allelochemicals) would mitigate decreased feeding deterrent response following prolonged exposure in this generalist herbivore. Neonate larvae were reared on cabbage leaves treated with individual feeding deterrents (digitoxin, thymol, toosendanin or xanthotoxin), or with binary mixtures of these until the third instar. Feeding deterrent responses to each antifeedant or mixture was then determined in a leaf disc choice bioassay. All of the mixtures produced additive deterrence when presented to naïve larvae. Larvae reared on individual antifeedants showed a significantly decreased feeding deterrent response (except to digitoxin), whereas larvae reared on binary mixtures of antifeedants did not show a decreased feeding deterrent response to any of them. Such mixtures were synergistic in terms of their feeding deterrence to experienced larvae. Our experiment supports the hypothesis (Jermy 1986) that mixtures of deterrents can prevent decreased feeding deterrent response following prolonged exposure, and provides one explanation for the multiplicity of chemical defenses found in many plants.     

Fungal pathogenesis of the sea fan Gorgonia ventalina: direct and indirect consequences

Summary. Fungal pathogenesis of the sea fan Gorgonia ventalina has developed on the Belize barrier reef during 1997 and 1998. The disease incidence (= percent of infected sea fans) remained unchanged at two sites along the barrier reef crest between years. However, the incidence increased significantly at an offshore atoll during that time, as did the virulence (= percent tissue loss). Grazing by the gorgonian specialist mollusc Cyphoma gibbosum increased on infected sea fans. Sea fan responses to fungal infection included significant decreases in a furano-germacrene compound with antifeedant activity and significant increases in the concentration of sclerites at the site of infection. Feeding assays utilizing C. gibbosum and a natural assemblage of reef fishes indicated sclerites are an effective deterrent, to both consumers, at both pre- and post-infected concentrations. In contrast, the compound was only deterrent to the fish, and only at concentrations found in pre-infected sea fans. These data indicate that sea fan pathogenesis can directly and indirectly affect population health via virulent tissue necrosis and pathogen-mediated reductions in grazer resistance, respectively. Received 4 November 1998; accepted 5 April 1999.     

Elysia timida (Risso, 1818) (Gastropoda, Opisthobranchia): relationship and feeding deterrence to a potential predator on the south-western Mediterranean coast

The relationship between the sacoglossan Elysia timida and the ornate wrasse Thalassoma pavo was studied in two laboratory experiments using artificial models. A feeding-preference experiment was conducted to determine whether mollusc extract deterred feeding by T. pavo, by using a "realistic" mollusc model (imitating the colour pattern of E. timida) coated with mollusc extract, and a reference model (without imitation or extract), and fishes collected from two locations. It was observed that fish approached, attacked and ate more reference models than mollusc models. A second feeding-preference experiment was designed with four different coloured models: "realistic" (W), green (G), red (R) and chequered (S) pattern. Both fish populations clearly rejected the S model, and differed in their colour preferences. Although both populations seemed to prefer the R and G models equally, the W model was clearly rejected by the fish that coexist with the mollusc at one site (Mazarrón), but was not rejected by the other population of fish which does not coexist with it (Cabo de Palos). Mazarrón fishes would identify the W model with the presence of a toxic compound during their coexistence, and therefore avoid attacking conspicuously coloured E. timida models as a response to their visual signals. Therefore, it was concluded that extract of E. timida is a deterrent for T. pavo, and its effect is sufficiently noxious that the fish tend to avoid it, so that the ability of the fishes to learn to recognise colours and identify certain colour patterns associated with obnoxious prey provides the molluscs the opportunity to survive by exhibiting a conspicuous coloration.     

Feeding deterrent and toxicity effects of apo-fucoxanthinoids and phycotoxins on a marine copepod (Tigriopus californicus)

Using the marine harpacticoid copepod Tigriopus californicus, the effects of phytoplankton feeding deterrents and toxins were differentiated and measured. Eight compounds were tested for feeding deterrence and toxicity responses: four apo-fucoxanthinoids (apo-10′-fucoxanthinal, apo-12′-fucoxanthinal, apo-12-fucoxanthinal, and apo-13′-fucoxanthinone) and four well-known phycotoxins (domoic acid, okadaic acid, microcystin-LR, and a mixture of PSP-1 toxins). Since several of these compounds exhibited both feeding deterrence and toxicity, a model was developed to deconvolute the observed toxicity response from the observed feeding deterrence response, and to classify these compounds based on the degree of toxicity and/or feeding deterrence they exhibited towards T. californicus. Microcystin-LR, the PSP-1 toxins, and the four apo-fucoxanthinoids behaved only as feeding deterrents at low concentrations. Okadaic acid exhibited both toxicity and feeding deterrence at low concentrations, with the threshold concentration for feeding deterrence at a lower level than the threshold concentration for toxicity. Domoic acid acted only as a toxin at low concentrations, with all decreases in feeding resulting from the death of the copepod. Received: 4 November 1996 / Accepted: 3 December 1996     

Color choices by bumble bees (Bombus terrestris): innate preferences and generalization after learning

It is usually assumed that the choice behavior of bees for floral colors is influenced by innate preferences only for the first flower visits prior to any experience. After visits to rewarding flowers bees learn to associate their colors with a reward. This learning process leads to an acquired preference for the trained colors that has been believed to dominate over previous experiences and over innate preferences. This work investigates how bumble bees (Bombus terrestris) chose among artificial flowers of different colors after they had been extensively trained to other colors. The bees chose novel colors according to their similarity to the trained color if the trained color was similar to some of the test colors. This was true also if trained colors and test colors were well distinguished, so their color choice reflected generalization between colors. If the test colors were so different from the trained color that no generalization took place, choice behavior was not affected by the trained color and reflected innate preferences. The differences in choice frequencies could not be explained by physical properties of the test colors other than the dominant wavelength, a parameter taken to reflect hue perception. Preferred dominant wavelengths correspond to those observed in naive bumble bees and honeybees. Thus bumble bees show innate preferences for certain colors not only prior to color learning but also after intensive learning when choosing among very different novel colors. Color choice among similar colors, however, is controlled by generalization from the learned color. Received: 9 November 1999 / Received in revised form: 19 March 2000 / Accepted: 31 March 2000     

Lingual and biting responses to prey chemicals by ingestively naive scincid lizards: discrimination from control chemicals,time course,and effect of method of stimulus presentation

Summary. We tested responses to prey chemicals by lizard hatchlings of an oviparous species and neonates of a viviparous species, neither of which had never eaten. Both species responded more strongly to prey chemicals than to odorous and odorless control stimuli presented on cotton swabs. Although only a few species have been examined, all that have been tested have an innate capacity for prey chemical discrimination, suggesting that this innate response to prey chemicals is widespread among lizards that use the lingual-vomeronasal system to locate and identify prey. Innate prey chemical discrimination has the great advantage of permitting lizards lacking prior experience with food to respond appropriately to chemical cues associated with food. Both species discriminated prey chemicals from control substances at age three days, earlier than previously known. Our data hint that Mabuya macularia may be capable of discrimination on its day of birth, but further study is needed to determine the exact onset. A stronger tendency to attack swabs bearing prey chemicals by Scincella lateralis than by M. macularia may be explained by differences in defensiveness near an experimenter or by differences in the importance of visual prey cues for confirmation of chemical cues in the natural habitats of these species. In M. macularia responses to the control stimuli declined over days of testing, suggesting habituation, but responses to prey chemicals did not habituate by the third day of testing, which is interpreted as a possible adaptive response to permit location of food. In the standard method of stimulus presentation, a cotton swab bearing a chemical stimulus is placed anterior to a lizard's snout. We tested a new method in which the swab was placed in continuous contact with the lizard's anterior labial scales. The new method elicited significantly stronger responses from M. macularia. We discuss reasons for this finding and applications for the new method. Received 2 September 1999; accepted 15 December 1999     

Effectiveness of tiger moth (Lepidoptera, Arctiidae) chemical defenses against an insectivorous bat (Eptesicus fuscus)

Summary. Adult tiger moths exhibit a wide range of palatabilities to the insectivorous big brown bat Eptesicus fuscus. Much of this variation is due to plant allelochemics ingested and sequestered from their larval food. By using a comparative approach involving 15 species from six tribes and two subfamilies of the Arctiidae we have shown that tiger moths feeding on cardiac glycoside-containing plants often contain highly effective natural feeding deterrents. Feeding on pyrrolizidine alkaloid-containing plants is also an effective deterrent to predation by bats but less so than feeding on plants rich in cardiac glycosides. Moths feeding on plants containing iridoid glycosides and/or moths likely to contain biogenic amines were the least deterrent. By manipulating the diet of several tiger moth species we were able to adjust their degree of palatability and link it to the levels of cardiac glycosides or pyrrolizidine alkaloids in their food. We argue that intense selective pressure provided by vertebrate predators including bats has driven the tiger moths to sequester more and more potent deterrents against them and to acquire a suite of morphology characteristics and behaviors that advertise their noxious taste.     

Lignoid chemical defenses in the freshwater macrophyte Saururus cernuus

Summary. Chemical defense against herbivores has rarely been investigated for freshwater plants, possibly due to the common misconception that herbivory on aquatic macrophytes is low and would not select for chemical defenses. In previous work, the freshwater angiosperm Saururus cernuus was shown to be a low preference food for omnivorous crayfish despite its high nutrient value and relatively soft texture. We used feeding by the crayfish Procambarus clarkii to guide fractionation of the deterrent lipid-soluble extract of this plant, leading to the identification of seven deterrent lignoid metabolites, (–)-licarin A, (+)-saucernetin, (–)-dihydroguaiaretic acid, (–)-sauriols A and B, (–)-saucerneol, and (–)-saucerneol methyl ether. Lignans have been implicated in terrestrial plant chemical defenses as insect growth inhibitors, insect toxins, nematocides, antibacterial, and antifungal agents. However, these activities have rarely been demonstrated using ecologically relevant methodologies in terrestrial systems, and never before in freshwater systems. The widespread nature of lignans amongst very distantly related plants, along with their rich diversity of molecular structure, suggests that they could play a large role in mediating plant-herbivore interactions. In addition to the lignoid compounds we identified, there were other compounds present in low concentration or unstable compounds that were deterrent, that did not appear to be lignans, but that we were unable to identify. This plant thus appears to be defended by a complex mixture of natural products. Received 6 June 2000; revised 23 August 2000; accepted 2 September 2000     

Mechanisms of benthic prey capture in wrasses (Labridae)

Teleost fishes capture prey using ram, suction, and biting behaviors. The relative use of these behaviors in feeding on midwater prey is well studied, but few attempts have been made to determine how benthic prey are captured. This issue was addressed in the wrasses (Labridae), a trophically diverse lineage of marine reef fishes that feed extensively on prey that take refuge in the benthos. Most species possess strong jaws with stout conical teeth that appear well-suited to gripping prey. Mechanisms of prey capture were evaluated in five species encompassing a diversity of feeding ecologies: Choerodon anchorago (Bloch, 1791), Coris gaimard (Quoy and Gaimard, 1824), Hologymnosus doliatus (Lacepède, 1801), Novaculichthys taeniourus (Lacepède, 1801) and Oxycheilinus digrammus (Lacepède, 1801). Prey capture sequences were filmed with high-speed video at the Lizard Island Field Station (14°40′S, 145°28′E) during April and May 1998. Recordings were made of feeding on pieces of prawn suspended in the midwater and similar pieces of prawn held in a clip that was fixed to the substratum. Variation was quantified among species and between prey types for kinematic variables describing the magnitude and timing of jaw, hyoid, and head motion. Species differed in prey capture kinematics with mean values of most variables ranging between two and four-fold among species and angular velocity of the opening jaw differing seven-fold. The kinematics of attached prey feeding could be differentiated from that of midwater captures on the basis of faster angular velocities of the jaws and smaller movements of cranial structures which were of shorter duration. All five species used ram and suction in combination during the capture of midwater prey. Surprisingly, ram and suction also dominated feedings on attached prey, with only one species making greater use of biting than suction to remove attached prey. These data suggest an important role for suction in the capture of benthic prey by wrasses. Trade-offs in skull design associated with suction and biting may be particularly relevant to understanding the evolution of feeding mechanisms in this group. Published online: 11 July 2002     

Ontogeny of diet changes in a tropical benthic carnivorous fish, Parupeneus barberinus (Mullidae): relationship between foraging behaviour, habitat use, jaw size, and prey selection

Parupeneus barberinus forages on benthic invertebrates using a wide range of foraging modes, including vigorous digging in the substratum, resulting in considerable disturbance to the benthos. Polychaetes were the most important prey item for all size classes, but fishes less than 120 mm total length consumed more small ostracods and nematodes than did larger fishes. Fishes greater than 120 mm total length consumed mostly bivalves, and fishes over 240 mm total length consumed mostly bivalves and crabs. A morphological examination of the feeding apparatus suggested that the size of important prey items consumed was determined by gape height and jaw width. Prey available to different size classes of fishes was determined by combining information on microhabitat use, foraging behaviours, and prey volumes in the substratum. Small fishes spent more time foraging on the reef flat and slope, compared with larger fishes that foraged mostly on the reef edge and base. In addition smaller fishes foraged mostly in the upper 2 cm of sediment, whereas larger fishes often foraged to depths of 10 cm. Selection ratios showed that different size classes of fishes selectively extracted different prey items from the substratum. Small fishes showed a preference for ostracods whereas large fishes selected for bivalves and crabs. Although polychaetes were the dominant prey item for all size classes, they were consistently selected against.     

Endemism and Regional Color and Genetic Differences in Five Putatively Cosmopolitan Reef Fishes

Abstract:  Endemism is thought to be relatively rare in marine systems due to the lack of allopatric barriers and the potential for long-distance colonization via pelagic larval dispersal. Although many species of coral reef fishes exhibit regionally restricted color variants that are suggestive of regional endemism, such variation is typically ascribed to intraspecific variation. We examined the genetic structure in 5 putatively monospecific fishes from the Indo-West Pacific ( Amphiprion melanopus , Chrysiptera talboti , and Pomacentrus moluccensis [Pomacentridae] and Cirrhilabrus punctatus , and Labroides dimidiatus [Labridae]) that express regional color variation unique to this area. Mitochondrial-control-region sequence analysis showed shallow to deep genetic divergence in all 5 species (sequence divergence 2–17%), with clades concordant with regional color variation. These results were partially supported by nuclear RAG2 data. An analysis of molecular variation (AMOVA) mirrored the phylogenetic results; Φ_ST values ranged from 0.91 to 0.7, indicating high levels of geographic partitioning of genetic variation. Concordance of genetics and phenotype demonstrate the genetic uniqueness of southwestern Pacific color variants, indicating that these populations are at a minimum distinct evolutionarily significant units and perhaps distinct regionally endemic species. Our results indicate that the alpha biodiversity of the southwestern Pacific is likely underestimated even in well-studied groups, such as reef fishes, and that regional endemism may be more common in tropical marine systems than previously thought.     

Effects of fish feeding by snorkellers on the density and size distribution of fishes in a Mediterranean marine protected area

Although there is a great deal of evidence to show that supplementary feeding by humans in terrestrial environments causes pronounced changes in the distribution and behaviour of wild animals, at present very little is known about the potential for such effects on marine fish. This study evaluated the consequences of feeding by snorkellers on fish assemblages in the no-take area of the Ustica Island marine protected area (MPA; western Mediterranean) by (1) determining if reef fish assemblage structure is affected in space and time by tourists feeding the fish; (2) assessing the effects of feeding on the abundance of the most common fish species; and (3) assessing the effects of feeding on the size structure of the two most numerically dominant ones. In particular, we hypothesised that both the abundance and the size structure of some fish species would increase at the study site following supplementary feeding, since the additional food provided by humans would make the site more appealing to them. Fish feeding influenced the fish assemblages within the Ustica MPA, and significant spatio-temporal changes occurred. While fish feeding appeared to have no effect on the ornate wrasse Thalassoma pavo, there was a noticeable increase in the number of Oblada melanura and Epinephelus marginatus in the impacted location after feeding. It is very likely that aggregations of fishes that evolve as a result of fish feeding by the public may have negative effects on local populations of fishes and invertebrates that make up their prey. Recreational use of coastal areas and MPAs is increasing elsewhere, making fish feeding a generalised human activity. Accurate information about its effect on the fish assemblage is essential to make responsible management decisions.Communicated by R. Cattaneo-Vietti, Genova     

Seaweed sex pheromones and their degradation products frequently suppress amphipod feeding but rarely suppress sea urchin feeding

Summary. A diverse group of brown seaweeds produce bouquets of C₁₁ metabolites, some of which act as pheromones that cue gamete release or attract sperm to eggs following release. We demonstrate that these C₁₁ metabolites and their degradation products also frequently and strongly deter feeding by the herbivorous amphipod Ampithoe longimana, but rarely by the herbivorous sea urchin Arbacia punctulata. Across the range of concentrations tested, seven of twelve C₁₁ metabolites or mixtures that we tested deterred feeding by the amphipod, but only two of eleven deterred the sea urchin. For those compounds where we could rigorously contrast the magnitude of deterrence against the amphipod with the magnitude of deterrence against the urchin, the amphipod was deterred significantly more than the urchin by five of six metabolites. Thus, C₁₁ compounds were more frequently and more strongly deterrent to the amphipod than to the sea urchin. These findings for C₁₁ metabolites conflict with previous investigations, where other classes of seaweed chemical defenses have been shown to deter feeding by large mobile herbivores like urchins and fishes but to be relatively ineffective against mesograzers, especially the species of amphipod that we used here. Our results suggest that C₁₁ metabolites are unusual among the known seaweed chemical defenses in that they are especially effective against mesograzers, which often consume seaweed spores, zygotes, and juveniles. The high concentrations of C₁₁ metabolites in brown algal eggs could allow these defenses to be especially important in defending gametes, zygotes, or young sporelings from herbivorous mesograzers. Received 26 February 1998; accepted 9 April 1998.     

Variability in the chemical defense of the sponge Chondrilla nucula against predatory reef fishes

Chondrilla nucula is a common Caribbean demosponge that grows in a range of habitats, from coral reefs to mangrove swamps. On reefs, C. nucula grows as a thinly encrusting sheet, while in mangrove habitats it surrounds submerged mangrove roots as fleshy, lobate clumps. Previous feeding experiments using predatory reef fish revealed a high degree of variability in the chemical defenses of C. nucula. The present study was undertaken to determine whether a relationship exists between habitat, growth form, and chemical defense of C. nucula. Both laboratory and field feeding-assays of crude extracts confirmed that C. nucula possesses a chemical defense with high intercolony variability, but there was no significant variation in feeding deterrency between reef and mangrove habitats at either geographic location (Bahamas and Florida). Extracts of C. nucula collected during September and October 1994 from the Bahamas were significantly more deterrent than those collected during August 1993, May 1994, and May 1995 from Florida, and extracts of these spring and summer Florida collections were more deterrent than extracts of C. nucula collected in December 1994 and February 1995 in the same locations. There was no evidence that deterrent compounds were concentrated in the surface tissues of the sponge, or that chemical defense could be induced by simulated predation. Laboratory and field assays of the fractionated crude extract revealed that feeding deterrency was confined to the most polar metabolites in the extract. Field transplants were used to determine whether predation influenced the growth form of C. nucula. Uncaged sponges transplanted from the mangrove to the reef were readily consumed by spongivorous reef fishes. Lobate mangrove sponges became thinner after being caged on the reef for 3 mo, but encrusting reef sponges did not become thicker after being caged in the mangroves for the same period of time. Reef sponges that were caged for 3 to 15 mo thickened by only a small amount (<1 mm) compared to uncaged and open-caged (i.e. in cages lacking tops) sponges. Simulated bite marks on both reef and mangrove sponges were repaired at a rapid rate (0.8 to 1.6 mm d⁻¹). Fish predation has an important impact on the distribution and abundance of C. nucula, but the thin growth form common to reef environments may be more the result of hydrodynamics than of grazing by spongivorous fishes. Received: 6 October 1997 / Accepted: 19 March 1998     

Social feeding in ladybird beetles: adaptive significance and mechanism

Summary. Young larvae of Adalia bipunctata search an area more intensively when exposed to the odour from other larvae feeding on aphids than when exposed only to the odour of aphids. In an olfactometer young larvae were significantly attracted either to the odour of crushed aphids or larvae feeding on aphids, but not to that of aphids, larvae, larvae plus aphids or larvae feeding on an artificial diet. That is, the change in searching behaviour appears in response to a volatile released by aphids when attacked. The odour released by crushed aphids is made up entirely of aphid alarm pheromone, β-farnesene. It is likely that the adaptive significance of this response is that it increases the ability of larvae to locate larvae that have already caught prey. By sharing the aphid kill of another larva it is likely that a first instar ladybird larva greatly increases its probability of surviving to the next instar. It is suggested that this social feeding is facilitated by egg clustering, which also may additionally account for why aphidophagous ladybirds lay their eggs in clusters. Received 28 February 2000; accepted 24 March 2000     

Seaweed secondary metabolites as antifoulants: effects of Dictyota spp. diterpenes on survivorship, settlement, and development of marine invertebrate larvae

Summary. A recent investigation showed that the brown seaweed Dictyota menstrualis was unfouled relative to co-occurring seaweeds, and that larvae of fouling invertebrates avoided settling on D. menstrualis due to chemicals on its surface. The secondary metabolites dictyol E and pachydictyol A are among the compounds found on this alga's surface. In the present study, we tested the effects of specific diterpenes from Dictyota on the survivorship, growth, and development of invertebrate larvae and developing juveniles that could foul seaweeds. Exposure to dictyol E, dictyol B acetate, pachydictyol A, and dictyodial from Dictyota menstrualis and D. ciliolata caused significant larval mortality, abnormal development, and reduce growth rates for three species of co-occurring invertebrates when their larvae were forced into contact with these metabolites. Larvae were damaged at metabolite concentrations as low as 5% of maximum possible surface concentrations of these compounds for the populations of Dictyota we studied. The negative effects of these secondary metabolites on potential foulers, in conjunction with data demonstrating larval avoidance of dictyol-covered surfaces, suggest that these compounds could function as chemical defenses against fouling, and could select for larvae that avoid hosts producing these metabolites. Received 25 May 1998; accepted 22 June 1998.     

Differences in the electroantennal responses of apple- and hawthorn-infesting races of Rhagoletis pomonella to host fruit volatile compounds

Summary. Domestic apple (Malus pumila)- and hawthorn (Crataegus sp.)-infesting races of Rhagoletis pomonella, Walsh (Diptera: Tephritidae) provide an excellent model to examine the role that host plant specificity plays during sympatric speciation (i.e., divergence in the absence of geographic isolation). Previous work has shown that these races differ in their propensities to accept apple and hawthorn fruits in behavioral choice assays, and that this discrimination translates into "host fidelity" in the field (i.e., apple flies tend to mate on and oviposit into apples and hawthorn flies on hawthorns). ?We present the results of a study examining possible physiological factors contributing to host choice differences in R. pomonella. We tested whether apple and hawthorn flies differ in their electroantennogram (EAG) responses to biologically relevant volatile compounds emitted from apples and hawthorns. Significant differences were found in the relative EAG responses of apple and hawthorn flies to host fruit compounds at five of six paired study sites across the eastern United States. The geographic pattern of EAG variation was complex, however, with local populations of apple and hawthorn flies tending to be more similar to one another than to flies of the same race at distant sites. This pattern was largely due to EAG responses for several compounds showing longitudinal or latitudinal clines, the latitudinal clines being similar to those observed for allozyme loci in the host races. We also found evidence for sex-related differences, as males tended to have higher mean EAG responses to compounds than females. Host-associated differences were therefore nested within geographic and sex-related differentiation in R. pomonella.?Further behavioral studies are needed to distinguish whether the EAG differences are responsible for, as opposed to being a consequence of, host-plant fidelity and adaptation. Crosses are also required to establish a genetic basis for the EAG responses, although we did find significant correlations between EAG scores for several compounds and the allozymes NADH-Diaphorase-2 and Hydroxyacid dehydrogenase at one of the study sites. Questions therefore remain concerning the evolutionary significance of the EAG response differences between apple and hawthorn fly races. Nevertheless, these differences raise the possibility that antennal responses to fruit-related volatile compounds contribute to host plant discrimination in R. pomonella. Regardless, the EAG responses represent another set of traits, in addition to diapause/eclosion time phenotypes and allozyme frequencies, differing between apple and hawthorn host races of R. pomonella. Received 17 March 1998; accepted 21 September 1989.     

Biofluorescence as a survey tool for cryptic marine species

As ecosystems come under increasing anthropogenic pressure, rare species face the highest risk of extinction. Paradoxically, data necessary to evaluate the conservation status of rare species are often lacking because of the challenges of detecting species with low abundance. One group of fishes subject to this undersampling bias are those with cryptic body patterns. Twenty‐one percent of cryptic fish species assessed for their extinction risk (International Union for Conservation of Nature [IUCN]) are data deficient. We developed a nondestructive method for surveying cryptically patterned marine fishes based on the presence of biofluorescence (underwater biofluorescence census, UBC). Blue LED torches were used to investigate how widespread biofluorescence was in cryptic reef fishes in the Coral Triangle region. The effectiveness of UBC to generate abundance data was tested on a data‐deficient pygmy seahorse species (Hippocampus bargibanti) and compared with data obtained from standard underwater visual census (UVC) surveys. We recorded 95 reef fish species displaying biofluorescence, 73 of which had not been previously described as biofluorescent. Of those fish with cryptic patterns, 87% were biofluorescent compared with 9% for noncryptic fishes. The probability of species displaying biofluorescence was 70.9 times greater for cryptic species than for noncryptic species. Almost twice the number of H. bargibanti was counted using the UBC compared with UVC. For 2 triplefin species (Ucla xenogrammus, Enneapterygius tutuilae), the abundance detected with UBC was triple that detected with UVC. The UBC method was effective at finding cryptic species that would otherwise be difficult to detect and thus will reduce interobserver variability inherent to UVC surveys. Biofluorescence is ubiquitous in cryptic fishes, making this method applicable across a wide range of species. Data collected using UBC could be used with multiple IUCN criteria to assess the extinction risk of cryptic species. Adopting this technique will enhance researchers’ ability to survey cryptic species and facilitate management and conservation of cryptic marine species.     

Artificial neural network modeling of caste odor discrimination based on cuticular hydrocarbons in termites

Summary. Individuals in an insect colony need to identify one another according to caste. Nothing is known about the sensory process allowing nestmates to discriminate minute variations in the cuticular hydrocarbon mixture. The purpose of this study was to attempt to model caste odors discrimination in four species of Reticulitermes termites for the first time by a non-linear mathematical approach using an "artificial neural network" (ANN). Several rounds of testing were carried out using 1 – the whole hydrocarbon mixtures 2 – mixtures containing the hydrocarbons selected by principal component analysis (PCA) as the most implicated in caste discrimination. Discrimination between worker and soldier castes was tested in all four species. For two species we tested discrimination of four castes (workers, soldiers, nymphs, neotenics). To test cuticular pattern similarity in two sibling species (R. santonensis and R. flavipes), we performed two experiments using one species for training and the other for query. Using whole hydrocarbons mixtures, worker/soldier discrimination was always successful in all species. Network performance decreased with the number of hydrocarbons used as inputs. Four-caste discrimination was less successful. In the experiment with the sibling species, the ANN was able to distinguish soldiers but not workers. The results of this study suggest that non-linear mathematical analysis is a good tool for classification of castes based on cuticular hydrocarbon mixture. In addition this study confirms that hydrocarbon mixtures observed are real chemical entities and constitute a true chemical signature or odor. Whole mixtures are not always necessary for discrimination. Received 23 July 1998; accepted 9 October 1998.     

Different phagostimulants in potato foliage for Manduca sexta and Leptinotarsa decemlineata

Summary. In the past decades, several studies have focused on the identification of feeding stimulants for specialists that feed on solanaceous plants, especially potato (Solanum tuberosum). In the 1950's, a phagostimulant was isolated from potato and tomato for Manduca sexta and characterised as a glycoside. It was suggested that the phagostimulant for M. sexta and Leptinotarsa decemlineata is identical. We tested whether these insects indeed share the same recognition factor. Previous bioassays for L. decemlineata larvae and adults were found to be unsatisfactory, so a new assay system was developed, using starch and wheat flour-based wafers as a neutral substrate. An aqueous extract of potato foliage that was highly active as a stimulant was fractionated by reverse phase medium pressure liquid chromatography (MPLC). Both insects were stimulated by a MPLC fraction that eluted with 25% methanol in water. Further separation of this and the following fraction (35% methanol, stimulatory for the beetle only) by semi-preparative high performance liquid chromatography (HPLC), using gradients of water and acetonitrile, resulted in the isolation of two distinct fractions that stimulated feeding by L. decemlineata. None of the HPLC fractions was active for M. sexta. However, fractions of the flash chromatography with less polarity (45–75% methanol) had a stimulatory effect only on M. sexta. Thus, the two insects do not use a common feeding stimulant, and for both at least two compounds of different polarity are active. Received 3 July 2000; accepted 24 October 2000